Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: UNIPROT:P06889 (Mol)
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The genome of Onchocerca volvulus was found to contain 2 actin gene classes (called 1 and 2) of 2 genes each. The 4 genes are located in 2 clusters (called A and B), each containing a gene class member. Five short introns of 122-207 bp occur within each gene. The sequences of the fourth intron and the 5' and 3' untranslated regions of all the 2 gene classes are completely different even though their coding regions share 95% identity. Mature transcripts from the actin genes have the nematode spliced leader (SL) at their 5' ends. One actin cDNA was found to be derived from an actin pre-mRNA which locks both the 5 introns and the 5' SL, suggesting that in at least some transcripts cis-splicing is completed before trans-splicing occurs.
Mol Biochem Parasitol 1992 Oct
PMID:The actin genes of Onchocerca volvulus. 143 69

We have isolated and characterized a gene encoding a novel GTP-binding protein of the GTPase superfamily in the filarial parasites Brugia malayi and Onchocerca volvulus. The deduced amino acid sequence of the cloned molecule has approximately 30% overall homology to ras proteins and approximately 90% homology to the 'ras-like' nuclear proteins TC4, ran and Spil. Rabbit antisera to bacterially expressed filarial protein detect a 24-22 kDa doublet in extracts of adult B. malayi and mature microfilariae, which is absent from immature microfilariae. Increased expression of the native parasite protein occurs when worms are cultured in the presence of epidermal growth factor.
Mol Biochem Parasitol 1992 Dec
PMID:Filarial parasites contain a ras homolog of the TC4/ran/Spil family. 148 50

The identification and characterization of a recombinant cDNA clone (OV103) expressing a microfilarial surface-associated antigen of Onchocerca volvulus is described. OV103 was identified and isolated from a lambda gt11 cDNA expression library derived from adult O. volvulus mRNA using a chimpanzee antiserum, taken 2 years after infection with third-stage larvae of O. volvulus. The cDNA clone encodes a 12.5-kDa protein that corresponds to a 15-kDa parasite protein present in microfilariae and adult female worms. The antigen encoded by this clone is located in the basal layer of the cuticle and the hypodermis of the female adult worm, and on the surface of microfilariae. OV103 fusion polypeptide is recognized only by some sera from onchocerciasis infected subjects (57%), but more significantly (89%) by sera from individuals that have low levels of patent infection. In addition, the antibody response to this protein developed before appearance of microfilariae in the skin of chimpanzees that had developed non-patent or low level patent infections, while the antibody response in chimpanzees with high levels of microfilariae appeared later at the time of appearance of microfilariae. Preliminary experiments indicated that affinity purified antibodies directed against OV103 fusion polypeptide mediated killing of nodular microfilariae in vitro in the presence of normal peripheral blood granulocytes.
Mol Biochem Parasitol 1992 Jan
PMID:Identification and characterization of an Onchocerca volvulus cDNA clone encoding a microfilarial surface-associated antigen. 154 18

Receptors potentially involved in neurotransmitting have been characterised in the muscle tissue and in whole worms of the nematodes Ascaris suum and Onchocerca volvulus, respectively. Binding studies revealed a high affinity for LSD with apparent KD values of 94 nM for A. suum and 120 nM for O. volvulus, whereas those of the neuroleptics haloperidol, spiperone and mianserin were found to be in the micromolar range. A variety of neurotransmitter antagonists, known to bind with high affinities either to mammalian D1/2 or to 5-HT1/2 receptors, were tested for their ability to bind to the nematode receptor. Results from these displacement experiments using tritiated LSD, mianserin, spiperone and haloperidol show distinct specificities of the nematode receptors compared to known receptor classes of mammals. With respect to this novel specificity, the nematode receptors seem to be unique and clearly distinct from those of the hosts.
Mol Biochem Parasitol 1991 Apr
PMID:Hallucinogenic and neuroleptic drug interactions with potential neurotransmitter receptors in parasitic nematodes. 167 21

The primary structure of an immunodominant antigen of the filarial parasite, Onchocerca volvulus was deduced from cDNA sequence analysis. Using affinity-purified antibody from onchocerciasis patients from West Africa, we have isolated a cDNA clone from a lambda gt11 cDNA expression library derived from microfilariae-producing female O. volvulus. The open reading frame encodes 152 amino acids, and the deduced sequence predicts a Mr of 16,850 (consistent with the apparent Mr of 18,000 of the immunoprecipitated in vitro translated product). The primary translation product contains a putative signal peptide of 16 amino acids. The mRNA coding for this antigen has an estimated size of 950 nucleotides. Furthermore, immunoelectron microscopy established that the antigen encoded by this clone is present in the hypodermis, the cuticle, and in the uterus of the filarial worms. Since this antigen is recognized exclusively by sera from onchocerciasis patients, and not by other sera from patients infected by other filarial parasites, it may prove to be an especially valuable tool for improving the specific diagnosis of onchocerciasis.
Mol Biochem Parasitol 1990 Feb
PMID:Identification of an Onchocerca volvulus cDNA encoding a low-molecular-weight antigen uniquely recognized by onchocerciasis patient sera. 168 59

Genomic DNAs of the related parasitic nematodes Onchocerca volvulus and Dirofilariae immitis, and a cDNA library of O. volvulus, were examined for the presence of the 22-nucleotide spliced leader (SL) found at the 5' ends of 10 to 15% of the mRNAs in the free-living nematode Caenorhabditis elegans. As in C. elegans, genes for the SL RNA are linked to the repetitive 5S rRNA genes of O. volvulus and D. immitis, but unlike C. elegans, they are in the same orientation as the 5S rRNA genes within the repeat unit. In O. volvulus the SL sequence is also encoded at more than 30 additional genomic locations and occurs at interior sites within many transcripts. Sequence determinations of four different cDNAs of O. volvulus, each containing an internal copy of the SL within a conserved 25mer, and one corresponding genomic DNA clone indicate that this sequence is not trans spliced onto these RNAs, but is encoded within the genes. The RNAs of two of these cDNAs appear to be developmentally regulated, since they occur in adult O. volvulus but were not detected in the infective L3 stage larvae. In contrast, actin mRNAs are present at all developmental stages, and at least one actin mRNA species contains a trans-spliced 5' SL. The internal locations of the SL in various transcripts and its perfect sequence conservation among parasitic and free-living nematodes argues that it serves specific, and perhaps multiple, functions for these organisms.
Mol Cell Biol 1990 Jun
PMID:Many transcribed regions of the Onchocerca volvulus genome contain the spliced leader sequence of Caenorhabditis elegans. 169 60

The complete coding sequence of Onchocerca volvulus myosin heavy chain has been determined from a series of overlapping cDNAs. The protein sequences from the 2 filarids, one responsible for subcutaneous filariasis, the other for lymphatic filariasis, show 92% identity, and are 1957 amino acids long. Each protein sequence is also equally related, with 75% identity, to MHC-B, the protein encoded by the unc-54 gene of the free-living nematode C.elegans. Such analysis is useful in phylogenetic studies among nematodes, as well as in structure-function relationships among myosin isolates.
Mol Biochem Parasitol 1992 Feb
PMID:Comparison of the body wall myosin heavy chain sequences from Onchocerca volvulus and Brugia malayi. 174 Oct 12

We report here the development of in situ hybridization and immunohistochemistry protocols which permit the histological identification of gene expression of a cloned antigen of Onchocerca volvulus, OI5, in the parasite. Skin nodules containing female adult worms were fixed in a modified Carnoy's fixative and embedded in paraffin. Histological staining of tissue sections revealed uniformly excellent morphology and RNA preservation. To localize mRNA by in situ hybridization, tissue sections were incubated with biotin-labeled pOI5, the plasmid containing the genomic sequence of the antigen, and hybridization signals were histochemically visualized using a streptavidin-enzyme conjugate and chromogenic substrates. The protein antigen was localized immunohistochemically by incubating the sections with specific antibodies prepared against a recombinant fusion protein containing the OI5 sequence (OI3), and visualized via a secondary antibody-biotin-enzyme conjugate procedure. The results reported here showed distinct localization of the OI5 mRNA and OI3 antigen in specific cellular and tissue regions of the adult parasite, and in microfilariae located within the uteri and in the surrounding host tissue. The specificity and high sensitivity of these histological detection methods should be generally applicable for the characterization of gene expression in the filarial parasite, particularly the insect-borne, infective filarial larvae, which are severely limited in quantity.
Mol Biochem Parasitol 1991 Dec
PMID:Histochemical localization of gene expression in Onchocerca volvulus: in situ DNA histohybridization and immunocytochemistry. 177 63

We have previously shown that an Onchocerca volvulus cDNA clone in lambda gt-11 designated OvG15, potentially encoding a peptide homologous to the 70-kDa heat shock protein (Hsp70), was recognized by sera of many individuals living in a zone endemic for lymphatic filariasis and most strikingly by sera from amicrofilaremic individuals including endemic normals, those with chronic symptoms and TPE patients. Few asymptomatic microfilaremics recognized the Hsp70. We have now used the insert from the OvG15 clone to isolate the homologous gene from Brugia malayi and analyze its primary structure and expression. The data presented in this communication describe a heat-inducible member of the hsp70 gene family of B. malayi which demonstrates intriguing features of tissue specific basal level expression, developmental regulation and heat inducibility.
Mol Biochem Parasitol 1991 Dec
PMID:Characterization of an hsp70 gene from the human filarial parasite, Brugia malayi (Nematoda). 177 66

We report here a panel of cDNA clones from Onchocerca volvulus which were isolated on the basis of being uniquely recognised by onchocerciasis sera and not by sera from patients infected with the major lymphatic filarial nematode parasite Wuchereria bancrofti. Over 90% of O. volvulus recombinants from a primary screen were found to cross-react with lymphatic filariasis sera and were discarded. The subset of specific clones, selected with pooled sera, was then screened with panels of individual patient sera. Individual onchocerciasis cases showed a highly heterogeneous pattern of recognition of recombinant peptides, but several clones were identified which could be combined in a cocktail of antigenic epitopes to successfully detect all infected cases in the study. All these clones encode low molecular weight proteins of the parasite, confirming earlier reports that antigens of this size class show greater species specificity. Several clones encode proteins of 20-23 kDa, the same molecular weight range as the major surface protein of adult worms. The two most commonly recognised clones, Ov22/31M and Ov20/36M were subcloned into the vector pNGS 8 which produces fusion proteins attached to a polyasparagine leader. The fusion peptides of both Ov22/31M and Ov20/36M were soluble and easily purified by gel filtration. Purified fusion protein was used in ELISA to assess reactivity of infected patients giving 90% sensitivity with 100% specificity.
Mol Biochem Parasitol 1991 Jun
PMID:cDNA clones of Onchocerca volvulus low molecular weight antigens provide immunologically specific diagnostic probes. 192 97


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